Search results
Results from the WOW.Com Content Network
Western blot workflow. The western blot (sometimes called the protein immunoblot), or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. [1]
Normalization of Western blot data is an analytical step that is performed to compare the relative abundance of a specific protein across the lanes of a blot or gel under diverse experimental treatments, or across tissues or developmental stages.
A dot blot (or slot blot) is a technique in molecular biology used to detect proteins. It represents a simplification of the western blot method, with the exception that the proteins to be detected are not first separated by electrophoresis. Instead, the sample is applied directly on a membrane in a single spot, and the blotting procedure is ...
Commercial DTNB may not be completely pure, so may require recrystallization to obtain completely accurate and reproducible results. [5] Ellman's reagent can be used for measuring low-molecular mass thiols such as glutathione in both pure solutions and biological samples, such as blood. [7] It can also measure the number of thiol groups on ...
Similar to a western blot, the far-western blot uses protein–protein interactions to detect the presence of a specific protein immobilized on a blotting matrix. Antibodies are then used to detect the presence of the protein–protein complex, making the Far-Western blot a specific case of the Western blot.
A sample extract of interest (for example human cells harvested from tonsil tissue) is placed in one well, sera or purified antibodies are placed in another well and the plate left for 48 hours to develop. During this time the antigens in the sample extract and the antibodies each diffuse out of their respective wells.
The blank solution should be the same pH and of a similar ionic strength as the sample solution. Example: using water for the blank measurement for samples dissolved in TE may result in low 260/230 ratios. A260/A280 Residual phenol or other reagent associated with the extraction protocol. A very low concentration (< 10 ng/μL) of nucleic acid.
Other interference may come from the buffer used when preparing the protein sample. A high concentration of buffer will cause an overestimated protein concentration due to depletion of free protons from the solution by conjugate base from the buffer. This will not be a problem if a low concentration of protein (subsequently the buffer) is used. [6]